To improve selectivity and specificity of cell membrane chromatography (CMC), the chromatography affinities of nine ligands of a1-adrenergic receptor(AR)to a1D-AR subtype were investigated. The human embryonic kidney ...To improve selectivity and specificity of cell membrane chromatography (CMC), the chromatography affinities of nine ligands of a1-adrenergic receptor(AR)to a1D-AR subtype were investigated. The human embryonic kidney (HEK) 293 cells expressed by cDNA of a1D-AR subtypes were cultured and cell membrane stationary phase (CMSP) was prepared. Then the interactions between ligands and a1D-AR in CMSP were investigated using CMC. The affinity rank order to a1D-AR subtype obtained from CMC for the nine a1-adrenoceceptor ligands is: prazosin, BMY7378, phentolamine, oxymetazoline, 5-methylurapidil, norepinephrine, phenyle- phrine, methoxamine, RS-17053. The affinity rank order is similar and correlates well with that obtained from others radioligand binding assays (RBA). CMSP prepared by transfected HEK293 cells with 1D-adrenoceptor cDNA and CMC method could be used to evaluate affinities of drug-receptor and drug-receptor subtypes and to screen drugs selective to a1D-AR.展开更多
Background Studies have confirmed that angiotensin II receptor blocker （ARB） and angiotensin converting enzyme inhibitors （ACEI） in the treatment of diabetic nephropathy （DN） has special advantages. We observed ...Background Studies have confirmed that angiotensin II receptor blocker （ARB） and angiotensin converting enzyme inhibitors （ACEI） in the treatment of diabetic nephropathy （DN） has special advantages. We observed the effects of valsartan and metoprolol tartrate hydrchloride in treatment of DN patients with positive β1-adrenergic and anti-angiotensin II type 1 （AT1） receptor antibody. Methods The epitopes of the second extracellular loop of β1 receptor （197-222） and AT1 receptor （165-191）, were synthesized and used respectively to screen serum autoantibodies from patients with DN （n=371, group A）, diabetes mellitus （DM） without renal failure （n=107, group B） and healthy blood donors （n=47, control, group C） by enzyme-linked immunosorbent assay （ELISA）. Metoprolol tartrate 25-50 mg, three times per day, valsartan 160 mg, once a day, aspirin 100 rag, once a day, and nitrendipine 10-20 mg, three times per day, were given to DN patients with positive or negative autoantibodies. The cystatin C level and 24-hour urinary protein were measured before and after treatment. Results In DN patients, the positive rate of the autoantibodies against β1 receptors and AT1 receptor was 47.7% and 51.5%, respectively, which were significantly higher than those in DM patients and healthy controls （all P 〈0.01）. Patients with anormalous cystatin C had higher positive rates of the autoantibodies than patients with normal cystatin C. Valsartan and metoprolol tartrate reduced proteinuria significantly （P 〈0.01） in DN patients with positive autoantibodies. Conclusions The findings suggest that these autoantibodies against β1 and ATl-receptor may play important roles in the pathogenesis of DN. Valsartan and metoDrolol tartrate are effective and safe in the treatment of DN.展开更多
Evidence suggests that the deterioration of communication between the sympathetic nervous system and cardiovascular system always accompanies the aging of human and animals. Cardiac sympathetic norepinephrine(NE) tran...Evidence suggests that the deterioration of communication between the sympathetic nervous system and cardiovascular system always accompanies the aging of human and animals. Cardiac sympathetic norepinephrine(NE) transporter(NET) on presynaptic membrane is a predominant component to eliminate released NE in the synaptic cleft and maintains the sensitivity of the β-adrenergic receptor(β-AR). In the present study,we investigated NET and β1-AR mRNA levels and sympathetic nerve density in cardiac sympathetic ganglion and left ventricular myocardium in 2-and 16-month-old rats with Northern blot analysis and immunohistochemistry. The expression levels of NET mRNA,NET protein and β1-AR mRNA in the ganglia or myocardia of 16-month-old rats were markedly reduced by 67%,26%,and 43%,respectively,in comparison with those in 2-month-old rats. Our results also show that aging induces a strong decrease of the catecholaminergic nerve fiber density.展开更多
目的研究比索洛尔对腹主动脉结扎所致慢性心力衰竭大鼠β_1肾上腺素能受体(β_1-AR)心功能的调节作用。方法 SD 大鼠40只,随机分为4组,即腹主动脉缩窄(COA)组;COA+比索洛尔组;假手术组;假手术+比索洛尔组。采用超声心动图和心室插管法...目的研究比索洛尔对腹主动脉结扎所致慢性心力衰竭大鼠β_1肾上腺素能受体(β_1-AR)心功能的调节作用。方法 SD 大鼠40只,随机分为4组,即腹主动脉缩窄(COA)组;COA+比索洛尔组;假手术组;假手术+比索洛尔组。采用超声心动图和心室插管法评估心功能,ELISA_法检测各组大鼠血浆去甲肾上腺素(NE)水平,RIA 法检测心肌腺苷酸环化酶(AC)活性,通过 RT-PCR 检测心肌β_1-AR、β肾上腺素能受体激酶1(β-ARK1)和 Gi 蛋白 mRNA 的表达变化,Western blot 检测β_1-AR 蛋白水平的表达变化。结果 COA 组大鼠心功能恶化,血浆 NE 水平明显升高,β_1-AR mRNA 表达减少、β-ARK1和 Gi 蛋白 mRNA 表达增加,β_1-AR 的蛋白水平表达减少,AC 活性明显下降;比索洛尔明显改善大鼠心功能[射血分数:COA+比索洛尔组:(66.7±8.4)%比 COA 组:(43.3±7.7)%,P<0.05],降低血浆 NE 水平[COA+比索洛尔组:(570.2±41.2)pg/mL 比 COA 组:(908.24±75.10)pg/mL,P<0.05],增高β_1-AR mRNA 表达[COA+比索洛尔组:(57.5±5.4)比 COA 组:(44.9±4.2)P<0.05],降低β-ARK1和 Gi 蛋白 mRNA 表达[β-ARK1:COA+比索洛尔组:(38.0±3.2)比 COA 组:(52.4±4.1),P<0.05;Gi 蛋白:COA+比索洛尔组:(72.8±5.8)%比 COA 组:(102.9±8.5)%,P<0.05],Western blot检测显示比索洛尔也增加β_1-AR 的蛋白水平表达[COA+比索洛尔组:(59.2±4.8)%比 COA 组:(30.9±2.3)%,P<0.05],AC 活性明显升高。结论比索洛尔能够调节慢性心力衰竭大鼠β_1-AR 信号转导通路,改善心功能,延缓心力衰竭进展。展开更多
文摘To improve selectivity and specificity of cell membrane chromatography (CMC), the chromatography affinities of nine ligands of a1-adrenergic receptor(AR)to a1D-AR subtype were investigated. The human embryonic kidney (HEK) 293 cells expressed by cDNA of a1D-AR subtypes were cultured and cell membrane stationary phase (CMSP) was prepared. Then the interactions between ligands and a1D-AR in CMSP were investigated using CMC. The affinity rank order to a1D-AR subtype obtained from CMC for the nine a1-adrenoceceptor ligands is: prazosin, BMY7378, phentolamine, oxymetazoline, 5-methylurapidil, norepinephrine, phenyle- phrine, methoxamine, RS-17053. The affinity rank order is similar and correlates well with that obtained from others radioligand binding assays (RBA). CMSP prepared by transfected HEK293 cells with 1D-adrenoceptor cDNA and CMC method could be used to evaluate affinities of drug-receptor and drug-receptor subtypes and to screen drugs selective to a1D-AR.
基金This study was supported by a grant from the Natural Science Foundation of Hubei Province （No. 2002AB 116）.
文摘Background Studies have confirmed that angiotensin II receptor blocker （ARB） and angiotensin converting enzyme inhibitors （ACEI） in the treatment of diabetic nephropathy （DN） has special advantages. We observed the effects of valsartan and metoprolol tartrate hydrchloride in treatment of DN patients with positive β1-adrenergic and anti-angiotensin II type 1 （AT1） receptor antibody. Methods The epitopes of the second extracellular loop of β1 receptor （197-222） and AT1 receptor （165-191）, were synthesized and used respectively to screen serum autoantibodies from patients with DN （n=371, group A）, diabetes mellitus （DM） without renal failure （n=107, group B） and healthy blood donors （n=47, control, group C） by enzyme-linked immunosorbent assay （ELISA）. Metoprolol tartrate 25-50 mg, three times per day, valsartan 160 mg, once a day, aspirin 100 rag, once a day, and nitrendipine 10-20 mg, three times per day, were given to DN patients with positive or negative autoantibodies. The cystatin C level and 24-hour urinary protein were measured before and after treatment. Results In DN patients, the positive rate of the autoantibodies against β1 receptors and AT1 receptor was 47.7% and 51.5%, respectively, which were significantly higher than those in DM patients and healthy controls （all P 〈0.01）. Patients with anormalous cystatin C had higher positive rates of the autoantibodies than patients with normal cystatin C. Valsartan and metoprolol tartrate reduced proteinuria significantly （P 〈0.01） in DN patients with positive autoantibodies. Conclusions The findings suggest that these autoantibodies against β1 and ATl-receptor may play important roles in the pathogenesis of DN. Valsartan and metoDrolol tartrate are effective and safe in the treatment of DN.
基金supported by the Postdoctoral Fellow Foundation of the Science and Technology Committee of Shanghai (No. 98-10)the Natural Science Foundation of Chinese People's Armed Police Force (Nos. WKH2006-5 and WKH2008ZO4), China
文摘Evidence suggests that the deterioration of communication between the sympathetic nervous system and cardiovascular system always accompanies the aging of human and animals. Cardiac sympathetic norepinephrine(NE) transporter(NET) on presynaptic membrane is a predominant component to eliminate released NE in the synaptic cleft and maintains the sensitivity of the β-adrenergic receptor(β-AR). In the present study,we investigated NET and β1-AR mRNA levels and sympathetic nerve density in cardiac sympathetic ganglion and left ventricular myocardium in 2-and 16-month-old rats with Northern blot analysis and immunohistochemistry. The expression levels of NET mRNA,NET protein and β1-AR mRNA in the ganglia or myocardia of 16-month-old rats were markedly reduced by 67%,26%,and 43%,respectively,in comparison with those in 2-month-old rats. Our results also show that aging induces a strong decrease of the catecholaminergic nerve fiber density.
文摘目的研究比索洛尔对腹主动脉结扎所致慢性心力衰竭大鼠β_1肾上腺素能受体(β_1-AR)心功能的调节作用。方法 SD 大鼠40只,随机分为4组,即腹主动脉缩窄(COA)组;COA+比索洛尔组;假手术组;假手术+比索洛尔组。采用超声心动图和心室插管法评估心功能,ELISA_法检测各组大鼠血浆去甲肾上腺素(NE)水平,RIA 法检测心肌腺苷酸环化酶(AC)活性,通过 RT-PCR 检测心肌β_1-AR、β肾上腺素能受体激酶1(β-ARK1)和 Gi 蛋白 mRNA 的表达变化,Western blot 检测β_1-AR 蛋白水平的表达变化。结果 COA 组大鼠心功能恶化,血浆 NE 水平明显升高,β_1-AR mRNA 表达减少、β-ARK1和 Gi 蛋白 mRNA 表达增加,β_1-AR 的蛋白水平表达减少,AC 活性明显下降;比索洛尔明显改善大鼠心功能[射血分数:COA+比索洛尔组:(66.7±8.4)%比 COA 组:(43.3±7.7)%,P<0.05],降低血浆 NE 水平[COA+比索洛尔组:(570.2±41.2)pg/mL 比 COA 组:(908.24±75.10)pg/mL,P<0.05],增高β_1-AR mRNA 表达[COA+比索洛尔组:(57.5±5.4)比 COA 组:(44.9±4.2)P<0.05],降低β-ARK1和 Gi 蛋白 mRNA 表达[β-ARK1:COA+比索洛尔组:(38.0±3.2)比 COA 组:(52.4±4.1),P<0.05;Gi 蛋白:COA+比索洛尔组:(72.8±5.8)%比 COA 组:(102.9±8.5)%,P<0.05],Western blot检测显示比索洛尔也增加β_1-AR 的蛋白水平表达[COA+比索洛尔组:(59.2±4.8)%比 COA 组:(30.9±2.3)%,P<0.05],AC 活性明显升高。结论比索洛尔能够调节慢性心力衰竭大鼠β_1-AR 信号转导通路,改善心功能,延缓心力衰竭进展。
